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PZ-FLIM-110
Piezo Scanning FLIM System
Based on bh’s Megapixel FLIM Technology Complete FLIM Microscopes FLIM Upgrades for Existing Microscopes Multidimensional TCSPC technique Sample Scanning by Piezo Stage Compact Electronics, Controlled by Laptop Computer Fully Integrated Scanner Control Confocal detection Suppression of out-of-focus light Suppression of lateral scattering Excellent contrast and resolution Excitation by bh BDL or BDS series picosecond diode laser Laser wavelengths 375nm, 405 nm, 445 nm, 473 nm, 488 nm, 515 nm, 640 nm, 685 nm, 785 nm Compatible with supercontinuum lasers 64-bit SPCM Software for Windows 7, 8, 10
Becker & Hickl GmbH Nahmitzer Damm 30 12277 Berlin, Germany Tel. +49 / 30 / 787 56 32 Fax. +49 / 30 / 787 57 34
email:
[email protected] www.becker-hickl.com
More than 22 years experience in multi-dimensional TCSPC. More than 1500 TCSPC systems worldwide. dbpzflim-02 Nov. 2015
PZ-FLIM-110
Piezo Scanning FLIM System
Megapixel FLIM Technology Record Images with up to 2048 x 2048 pixels Record FLIM data of a large number of cells under identical conditions Select regions of interest from recorded data of a large field of view
Lateral and Vertical Scanning
Select between a scan in the x-y plane or a scan in the x-z plane Run a vertical scan within a fraction of the time needed for a full Z stack
Phosphorescence Lifetime Imaging Simultaneously with FLIM Obtain unprecedented PLIM sensitivity by multi-pulse excitation Record phosphorescence and fluorescence lifetime images simultaneously
More than 22 years experience in multi-dimensional TCSPC. More than 1500 TCSPC systems worldwide.
dbpzflim-02 Nov. 2015
PZ-FLIM-110
Piezo Scanning FLIM System
Interactive Scanner Control Conveniently select scan field, pixel numbers, and scan speed Select a region of interest within a previously recorded image Record an image of the region selected
Predefined Setups Pass from one operation mode or system configuration to another by selecting from a list of ‘Predefined Setups’
Precision FLIM and PLIM Data Analysis Analyse FLIM and PLIM data pixel by pixel No need to record an IRF Up to three exponential decay components Calculate lifetimes and amplitudes of decay components Calculate ratios of lifetimes or amplitudes, fractional intensities, FRET efficiencies Calulate 1D and 2D histograms of decay parameters over the pixels Analyse large data volumes by batch processing
More than 22 years experience in multi-dimensional TCSPC. More than 1500 TCSPC systems worldwide.
dbpzflim-02 Nov. 2015
PZ-FLIM-110
Piezo Scanning FLIM System
System Components Laser High power High repetition rate Single-mode fibre coupling
BDL-SMN series
Repetition rates Available wavelengths Pulse width
BDS-SM Series
20 MHz, 50 MHz, 80 MHz, CW 20 MHz, 50 MHz, CW 375nm, 405 nm, 445 nm, 473 nm, 488 nm, 515nm, 640nm, 685nm, 785nm typ. 60 ps, depending on wavelength version and power
PZ FLIM Confocal Optics Assembly BDL-SMN or BDS series ps diode laser
Laser input Excitation / Emission Beamsplitter Emission Filter Output to detector Confocal pinhole
Microscope Side Port
Filter to Detector Fibre Adapter
Dichroic Mirror
Qioptiq Kineflex fibre manipulator Dichroic, different transition wavelength available In filter slider, long-pass and bandpass filters available Multi-mode fibre, FCS connector Core diameter of detector fibre
Microscope Lens
PZ FLIM Confocal Optics Assembly
Scan Stage Sample
Mad City Labs Nano View 200-3 1
Scan Stage
Detector
Wavelength Range Wavelength Channels TCSPC Module
Scan Control Module
Id 100-50-FC SPAD (Standard)
HPM-100-40 (optional)4
MW FLIM GaAsP (optional)4
400nm to 1000nm 1
400 nm to 700 nm 1
400 nm to 700 nm 16
SPC-150 (Standard)
SPC-150N (Optional)
GVD-120 Generation of scan signals Image size Laser multiplexing Laser on/off for PLIM Beam park function
SPC-160 (Optional)
Hardware, digital signal synthesis 16 x 16 to 2048 x 2048 pixels Inside pixel, line by line, or frame by frame2 Selectable in % of pixel time Any location within scan area
System Computer and Electronics Box Lap-top PC with PCI-extension box
Also available:
Extension box contains: SPC-150, SPC-150N, SPC-160 TCSPC modules GVD-120 scan controller DCC-100 detector controller (optional) SPC, GVD, and DCC modules installed in standard PC
More than 20 years experience in multi-dimensional TCSPC. More than 1500 TCSPC systems worldwide.
dbpzflim-02 Nov. 2015
PZ-FLIM-110
Piezo Scanning FLIM System
Specifications Scanning Optical principle Laser input Output to detector Main beamsplitter Pinhole Emission filter Connection to microscope Scan Controller Principle Scan waveform Scan format Frame size, frame scan Line scan X scan Y scan Laser power control, electrical Laser multiplexing 2 Beam blanking Scan rate Fastest scan rate with piezo stage 3 Scan area definition Beam park position Laser control Excitation Source Available Wavelengths Pulse width, typical Pulse frequency Power in picosecond mode Fast on/off modulation Detector (standard) Spectral Range Peak quantum efficiency IRF width with bh diode laser Background count rate, thermal Power supply Detector (optional) Spectral Range Cathode type Peak quantum efficiency IRF width with bh diode laser Background count rate, thermal Background from afterpulsing Power supply and overload shutdown Detector (optional) No. of wavelength channels Wavelength channel width Spectral Range Cathode type IRF width with bh diode laser Power supply and overload shutdown
Mad City Labs Nano-View 200-3 1 Sample scanning by piezo stage, confocal detection Single-mode fibre coupled, Qioptiq fibre coupling system Fibre coupling, FC connector, multi-mode fibre Dichroic Mirror Pinhole is core of detection fibre Filter slider adapter to left side port bh GVD-120 Digital waveform generation, scan waveforms generated by hardware [1] Linear ramp with cycloid flyback. Parameters configurable for different scanners line, frame, or single point 16x16 to 2048x2048 pixels 16 to 2048 pixels continuous or pixel-by-pixel line by line via electrical signal to lasers frame by frame, line by line, or within one pixel during flyback and when scan is stopped automatic selection of fastest possible rate or manual selection 5 lines / second via zoom and offset or interactive via image cursors selectable via cursor in FLIM image On/off, power, wavelength multiplexing 2 bh BDL-SMC Series or BDS-Series picosecond diode laser 375nm, 405nm, 445nm, 473nm, 488nm, 510nm, 640nm, 685nm, 785nm 40 to 70 ps BDL-Series: 20MHZ, 50MHz, 80MHz BDS Series: 20 MHZ, 50 MHz Typ. 0.2 mW to 2 mW injected into fibre. Depends on wavelength version < 1µs, for PLIM and excitation wavelength multiplexing2 Id Quantique id100-50-FC SPAD 400 to 1000 nm 50% 70 to 130 ps Typ. 100 to 200. LN version with <10 available 6V wall-mounted power supply bh HPM-100-40 hybrid detector [1] 4 400 to 700nm GaAsP 40 to 50% 120 to 130 ps 300 to 2000 counts per second not detectable via DCC-100 controller of TCSPC system bh Multi-Wavelength MW-FLIM Detector [1] 16 12.5 nm, other on requets 200 nm within a range of 400 to 700nm GaAsP 250 ps via DCC-100 controller of TCSPC system
4
More than 20 years experience in multi-dimensional TCSPC. More than 1500 TCSPC systems worldwide.
dbpzflim-02 Nov. 2015
PZ-FLIM-110
Piezo Scanning FLIM System
TCSPC System Principle Electrical timing jitter Electrical IRF width Minimum time channel width Dead time Saturated count rate Dual-time-base operation Source of macro time clock Input from detector Reference (SYNC) input Synchronisation with scanning Scan rate Synchronisation with laser multiplexing 2 Recording of multi-wavelength data 5 Basic acquisition principles, see [1]
bh SPC-150, SPC-150N, or SPC-160 module, see [1] for details Advanced TAC/ADC principle 2.3 ps rms 6.8 ps FWHM 813 fs 100 ns (80 ns for SPC-160) 10 MHz (12 MHz for SPC-160) via micro times from TAC and via macro time clock internal 40MHz clock or from laser constant-fraction discriminator constant-fraction discriminator via frame clock, line clock and pixel clock pulses any scan rate 3, automatic accumulation of frames via routing function simultaneous in all wavelength channels, via routing function on-board-buildup of photon distributions buildup of photon distributions in computer memory generation of parameter-tagged single-photon data online auto or cross correlation and PCH Operation modes, see [1] f(t), oscilloscope, f(txy), f(t,T), f(t) continuous flow FIFO (correlation / FCS / MCS) mode Scan Sync In imaging, Scan Sync In with continuous flow FIFO imaging, with MCS imaging, mosaic imaging 6, time-series imaging 6 Multi-wavelength operation 5, laser multiplexing operation 2 cycle and repeat function, autosave function Max. Image size, pixels (SPCM 64 bit software) 2048x2048 1024x1024 512x512 256x256 Max. no of time channels, see [1] 256 1024 4096 4096
SPCM Data Acquisition Software, please see [1] for details Windows 7, Windows 8, or Windows 10, 64 bit Operating system Loading of system configuration single click in predefined-setup panel Start / stop of measurement by operator or by timer, starts with start of scan, stops with end of current frame Online calculation and display, FLIM, PLIM in intervals of Display Time, min. 1 second Online calculation and display, FCS, PCH in intervals of Display Time, min. 1 second Number of images diplayed simultaneously max 8 Number of curves (Decay, FCS, PCH, Multiscaler) 8 in one curve window Cycle, repeat, autosave functions user-defined, used for for time-series recording 6, Z stack FLIM 6, microscope-controlled time series Saving of measurement data User command or autosave function Optional saving of parameter-tagged single-photon data Link to SPCImage data analysis automatically after end of measurement or by user command SPCImage FLIM and PLIM Data Analysis, please see [1] for details Data types FLIM, PLIM, single fluorescence or phosphorescence decay curves Decay models Single-, double-, triple-exponential decay models, incomplete decay models, first moment Procedure Iterative convolution or direct calculation by first moment Batch processing for multi-file FLIM data Parameters delivered by analysis lifetimes of components, amplitudes, ratios of amplitudes or lifetimes, FRET efficiency, fractional intensities of decay components Display Colour-coded images of decay parameters Histogram of decay parameters over pixels 2D histograms of decay parameters, phasor plot
Remarks 1. 2. 3. 4. 5. 6.
Other scan stages on request. Stage controller must have analog inputs. Laser multiplexing requires second laser and single-mode beam combiner. Limited by piezo stage. Fastest system scan rate depends on stage type. Configurable for different stages. Requires DCC-100 detector controller card. Requires MW FLIM detector. Limited applicability due to slow scan rate of piezo stage.
More than 20 years experience in multi-dimensional TCSPC. More than 1500 TCSPC systems worldwide. dbpzflim-02 Nov. 2015
PZ-FLIM-110
Piezo Scanning FLIM System
Literature (please contact bh for printed copies) The bh TCSPC Handbook, www.becker-hickl.com W. Becker, Advanced Time-correlated single photon counting techniques. Springer 2005 3. W. Becker (ed.), Advanced time-correlated single photon counting applications. Springer (2015) 4. The PZ-FLIM piezo-scanning FLIM system. Application note, available on www.becker-hickl.com 1. 2.
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International Sales Representatives
US: Boston Electronics Corp
[email protected] www.boselec.com
UK: Photonic Solutions
[email protected] www.photonicsolutions.co.uk
Japan: Tokyo Instruments Inc.
[email protected]. jp www.tokyoinst.co.jp
dbpzflim-02 Nov. 2015
China: DynaSense Photonics Co. Ltd.
[email protected] www.dyna-sense.com
